Focal cerebral ischemia in the TNFalpha-transgenic rat.
BACKGROUND
To determine if chronic elevation of the inflammatory cytokine, tumor necrosis factor-alpha (TNFalpha), will affect infarct volume or cortical perfusion after focal cerebral ischemia.


METHODS
Transgenic (TNFalpha-Tg) rats overexpressing the murine TNFalpha gene in brain were prepared by injection of mouse DNA into rat oocytes. Brain levels of TNFalpha mRNA and protein were measured and compared between TNFalpha-Tg and non-transgenic (non-Tg) littermates. Mean infarct volume was calculated 24 hours or 7 days after one hour of reversible middle cerebral artery occlusion (MCAO). Cortical perfusion was monitored by laser-Doppler flowmetry (LDF) during MCAO. Cortical vascular density was quantified by stereology. Post-ischemic cell death was assessed by immunohistochemistry and regional measurement of caspase-3 activity or DNA fragmentation. Unpaired t tests or analysis of variance with post hoc tests were used for comparison of group means.


RESULTS
In TNFalpha-Tg rat brain, the aggregate mouse and rat TNFalpha mRNA level was fourfold higher than in non-Tg littermates and the corresponding TNFalpha protein level was increased fivefold (p <or= 0.01). Infarct volume was greater in TNFalpha-Tg rats than in non-Tg controls at 24 hours (p <or= 0.05) and 7 days (p <or= 0.01). Within the first 10 minutes of MCAO, cortical perfusion measured by LDF was reduced in TNFalpha-Tg rats (p <or= 0.05). However, regional vascular density was equivalent between TNFalpha-Tg and non-Tg animals (p = NS). Neural cellular apoptosis was increased in transgenic animals as shown by elevated caspase-3 activity (p <or= 0.05) and DNA fragmentation (p <or= 0.001) at 24 hours.


CONCLUSIONS
Chronic elevation of TNFalpha protein in brain increases susceptibility to ischemic injury but has no effect on vascular density. TNFalpha-Tg animals are more susceptible to apoptotic cell death after MCAO than are non-Tg animals. We conclude that the TNFalpha-Tg rat is a valuable new tool for the study of cytokine-mediated ischemic brain injury.
Pettigrew
L Creed
LC
Sanders-Brown Center on Aging, University of Kentucky, Lexington, Kentucky, USA. cpetti@uky.edu
Kindy
Mark S
MS
Scheff
Stephen
S
Springer
Joe E
JE
Kryscio
Richard J
RJ
Li
Yizhao
Y
Grass
David S
DS
D000818:Animals; D017209:Apoptosis / Q000235:genetics / Q000276:immunology; D001921:Brain / Q000276:immunology* / Q000473:pathology / Q000503:physiopathology; D020520:Brain Infarction / Q000276:immunology* / Q000473:pathology / Q000503:physiopathology; D002545:Brain Ischemia / Q000276:immunology* / Q000503:physiopathology; D053148:Caspase 3 / Q000378:metabolism; D002536:Cerebral Arteries / Q000276:immunology / Q000473:pathology / Q000503:physiopathology; D004195:Disease Models, Animal; D004660:Encephalitis / Q000276:immunology* / Q000473:pathology / Q000503:physiopathology; D005260:Female; D020244:Infarction, Middle Cerebral Artery / Q000276:immunology / Q000503:physiopathology; D008297:Male; D008833:Microcirculation / Q000235:genetics / Q000276:immunology; D009474:Neurons / Q000276:immunology / Q000473:pathology; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D055647:Rats, Transgenic; D012039:Regional Blood Flow / Q000235:genetics / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000276:immunology*; D015854:Up-Regulation / Q000235:genetics / Q000276:immunology*
D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D053148:Caspase 3